Journal of Biomedicine the Future of Forensic Biology

Journal of Biomedicine 2018, Vol. 3 13

Ivyspring International Publisher Journal of Biomedicine 2018; 3: 13-18. doi: 10.7150/jbm.22760 Re view The Future of Forensic Biology Ajay Kumar Rana

Division of Biology, State Forensic Science Laboratory, Ministry of Home Affairs, Government of Jharkhand, Near Birsa Munda Jail, Hotwar, Ranchi, Jharkhand, India- 835217.

 Corresponding author: Tel.: +91-8292417706; Fax: +91-0651-2562557. Email: [email protected]

© Ivyspring International Publisher. This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions.

Received: 2017.09.09; Accepted: 2017.12.02; Published: 2018.01.23

Abstract The recent technological and scientific development in the field of forensic biology is certainly stamping a huge impact in capturing and convicting the criminals without any doubt. This review enlists forthcoming emerging technologies in forensic biology which would become inevitable and routine tasks in laboratories for solving extremely challenging and critical cases. These include touch DNA profiling, forensic phenotyping, 3D facial reconstruction and morphometrics, microbial forensics, virtual autopsy, DNA methylation and next-generation sequencing methods which would prove boon sooner or later for the crime investigators in finding missing persons, nabbing murderers and sexual assaulters and solving an array of cold cases.

Key words: Next generation forensic biology, Touch DNA, Forensic phenotyping, Morphometrics, Microbial forensics, Virtopsy, DNA methylation

Introduction The forensic science today has been expanded resolve many complex cases [15, 16]. Advances in with numerous innovative technological advances forensic biology, through the introduction of next and molecular discoveries. Following refinement and generation systems in photographical, biological, rigorous testing, many methods/technologies have serological and genetic analyses, are the only ways been adopted by forensic laboratories, including through which more direct, indirect, corroborative or polymerase chain reaction (PCR) [1, 2], capillary additive evidences can be collected to be presented in electrophoretic instrumentation (Genetic Analyzers) the court of law. The forensic laboratories are [3, 4], automated liquid handling (Microfluidic expected to migrate in near future from today’s devices) [5-7], and expert software systems [8-11]. current methods to more advanced and efficient Introduction of robust validation processes has a long technological platforms (Figure 1). In addition, term impact to test evidentiary samples to be forensic facilities that offer specialized testing for presented to the court of law. For example, the missing persons and mass disaster victim credibility of DNA has resulted in DNA becoming the identification will gain great capabilities leveraging gold standard for forensics and the most effective these next generation testing systems. Here is the list crime fighting tool available to law enforcement [12]. of advancing technologies that proffer to solve many The progression from Sanger-type sequencing inapprehensible, recalcitrant and critical cases (STS) using dideoxynucleotide triphosphates without any doubt. (ddNTPs) to next generation sequencing (NGS) has further made an impact in the field of forensic science, Beyond fingerprint Analysis other than in human health areas such as regenerative The fingerprints/footprints primarily consist of or personalized medicine [13, 14]. Today, sequencing organic/inorganic materials [17] or sometimes fats of mitochondrial (mtDNA) is already a validated, secreted from the skin which could reveal a horde of widely accepted and utilized tool which is helping to information about the persons who left them. In many instances, some skin cells could be dislodged on the

http://www.jbiomed.com Journal of Biomedicine 2018, Vol. 3 14 object while being handled as such or touched with ploning through φ-29 polymerase [21]. These any part of the body. In the future, fingerprints’ DNA technologies which helps to fish out touch DNA are (touch DNA) from these leftover skin cells on various therefore potentially promising to identify the traces surfaces can be analyzed through atomic force biological evidence and could be used to develop of microscope [18]or even can be collected through genetic profiles from incomprehensible/single cell optical tweezers equipped with SteREO Lumar.V12 human samples (theoretically as well as practically). stereomicroscope and UV unit [19], followed by This will allow the crime investigators to identify and amplification either through AmpFlSTR® NGM™ correctly nab the culprit in the court of law. Amplification Kit (Applied Biosystems) [20] or

Figure 1. The Next Generation Technologies/Methods in Forensic Biology. A. Morphometric analysis of skull and photographs using next generation 3-D facial reconstruction software using sophisticated laptops. B. Prediction of the color of skin, hair as well iris by sequencing/identifying the genes that actually code for the particular trait/color. C. Study of microbes from human body especially from pubic hair after sexual assault which establishes the exchange of microbial flora in between them. D. Virtual autopsy (Virtopsy®) is a non-invasive method to collect the images of the body through µCT and µMRI to be used in future when may be further required in medico-legal procedures. E. Touch DNA is trace amount of foreign DNA recovered from the person’s body during the course of hard press, strangling or kiss from another person. F. DNA methylation analysis is a new method being developed to predict the age of persons, discern monozygotic twins as well as identify various biological fluids recovered from scene of crime. G. Next generation serology involves the use of mass spectrometry especially tandem MS (protein sequencing) which will reveal the origin of the source (Human vs. Animal) through matching the obtained sequence with the non-redundant protein databases, such as UNIPROT. H. Microfluidic chips (for purification, amplification and capillary electrophoresis of DNA) have been developed which are so handy and time efficient that DNA profile of the source of biological exhibit can be established at the scene of crime within a day.

http://www.jbiomed.com Journal of Biomedicine 2018, Vol. 3 15

From Genotyping to Forensic Phenotyping modify a 3D modelled person’s attitude or position! For smartphones, the algorithms must be scaled Human phenotype such as hair color, iris color down, which makes them less effective, but in future and skin color can be predicted with the type of single with more sophistication of mobile phones it may nucleotide polymorphism (SNPs) harbored by coding become a handy tool for police officers/forensic (SLC45A2, SLC24A5, and MC1R)) as well as some scientists to recognize the accused/victim from a non-coding (HERC2 and ASIP) genes in the human crowd [28]. A recent report has analyzed whole genome [22]. Alleles of other pigmentation genes such genomic sequencing data from different ethnic as TYRP1, DCT and many more have been statistically community around the world (#1061) which is able to associated with human iris pigmentation or other identify people according to their physical traits and human pigmentation. These SNPs data have been demography [29]. Morphometrics (the measurement of translated by a biotech company (Thermo Fisher body shapes) may be used to identify the skeletal Scientific) to develop a forensic phenoytyping system remains of missing children, something which is called HIrisPlex which employs 24 DNA variants that extremely challenging for current forensic experts. helps to predict eye and hair color along with six Currently, the most commonly used form of genetic markers which helps to predict blonde hair morphometric methods are Generalized Procrustes 69.5 percent of the time, brown hair 78.5 percent of the method (GPA), Euclidean Distance Matrix Analysis time, red hair 80 percent of the time, and black hair (EDMA), Principal Components Analysis (PCA) and 87.5 percent of the time [23-25]. This system is based 3D imaging methods (µMRI and µCT) [30]. One of the on single base extension of primers using a fluorescently major aspect of morphometrics is allometric labelled ddNTP terminator ( SNaPshotTM chemistry) in measurements which compares size of the body parts developing polymorphic SNPs profiles of individuals in relation to the size of the organism [31]. Allometry using Genetic Analyzer (Applied Biosystems, Foster is also a prominent feature of contour variation for City, CA ) [25]. After inclusion of human skin color multifarious morphological traits such as craniofacial variant alleles (SNPs), this phenotyping system is shape [30]. palpably going to become an important instrument for routine investigation of questioned human color in Microbial Forensics the near future of forensics. The most recent report on the ratio of Facial Recognition Software and microbiome (largely the bacteria present in our body) to the number of cells in human body is roughly 4:3, Morphometrics of the same order (x1013), and the microbes Automated facial recognition software can be approximately weighs as 0.2 kg in a typical 70 kg used to identify a victim in disaster or match an human body [32]. Microbial forensics is a recent accused to a police database [26, 27]. If a photo discipline in the field of law which employs database of good quality exists such as Interstate microbiological methods for analysis of evidence in Photo System (IPS) possessed by FBI’s Next assorted forms of crimes which includes bioterrorism, Generation Identification System (NIS) in a utopian hoaxes, accidental release of biological agent/toxin or police system, then by using a sophisticated laptop even rape cases [33-35]. The vast microbial one can search a victim/suspect with facial communities (bacteria, yeast and viruses) remain recognition software among millions, barring the same except after sex. Therefore in future, the pubic stipulation that people face do not change much over hair recovered from sexual assault suspects if do not time. These softwares are typically used in security contain the roots, then the metagenomic analysis of systems (biometrics) and can be had from various microbes in the hair may become a substitute of sources such as evidence [36]. The microbes and hence the microbial • http://www.bioenabletech.com/tag/free-face-r DNA differs in males and females and therefore after ecognition-software/ sex, the microbes in both the male and the female • https://www.facefirst.com/ appear to swap, indicating that a sexual act occurred between a particular man and woman. The microbial • https://facedetection.com/software/ forensics has been broadened by metagenomic These companies have developed software that analysis of the microbes from the surroundings also recognizes 3D picture of a person’s face” using 3D such as from soil [37] in case of sensors that identifies distinctive features such as bioterroism/murderous crimes, which may also help globular cranial vault, mandible bone shape, eye in collecting and corroborating evidences to correlate sockets and nasal bone. The software’s users can also with the incident.

http://www.jbiomed.com Journal of Biomedicine 2018, Vol. 3 16

Virtual Autopsy (Virtopsy) [55] would be more assertively examined by Now the human dead body can be preserved proteomic analysis of characteristic markers present virtually for an infinite time which can help a in bodily fluid or tissues (hard or soft) viz. medico-legal doctor/scientist to examine the body by hemoglobin (blood), α-amylase 1 (Saliva), cornulin, conducting virtual autopsies even in its absence when cornifin, and/or involucrin (vaginal fluid), for religious, personal, or other reasons, spouses or semenogelin, prostate-specific antigen, acid families sometimes do not want their murdered loved phosphatase (semen), Uromodulin (urine), plunc one’s remains to be autopsied [38]. Invented and protein (nasal secretions), and immunoglobulins trademarked by Prof. Richard Dirnhofer, the virtopsy without hemoglobin, stercobilin (faeces). The are noninvasive, damaging neither the body nor sequence of protein derived from tandem MS forensic evidence [39, 40]. Virtopsy employs (MS-MS) would help to identify human as well as post-mortem computed tomography (PMCT), and is non-human tissues/secretions that could be easily best for gas embolism, pneumothorax pulmonary searched in freely available non-redundant database emphysema and the relationship between the internal of protein such as UNIPROT [56]. path of the instrument of aggression and the entry wound. It is quite expensive, but the cost is expected Conclusive Remarks and Future to decrease in coming near future as virtual autopsies Prospective will become rampant in forensics especially in For forensically questioned samples, unswerving medico-legal arena. tests and time efficient results can be only expected from laboratories when it is immediately processed

DNA methylation using standard protocols and advanced sophisticated The field of epigenetics has recently made a instruments. Therefore it presumably summons to call landmark entry in the field of forensic sciences for the the modernized laboratory very close to the crime identification and analysis of forensic samples which scene. The so called portable forensic labs which may otherwise contain exactly the same DNA sequences require handheld electronic sniffers and microfluidic [64]. DNA methylation particularly 5mC devices in the toxicology or DNA labs respectively, (5-methylcytosine) methylation at the CpG sites, the may allow scientists to conduct the experiment at the major epigenetic and chemical change in the DNA very crime scene and present instant results to the [41] which without altering the DNA sequence leads relevant court of law. The DNA technologies to phenotypic variation among the various tissues, combined with next-generation sequencing methods monozygotic twins or among humans has provided a will certainly save the time and deliver accuracy in new vista to distinguish cell types or tissues [42, 43], analyzing genomic data obtained from tissues [13, 57]. monozygotic twins [44-46], age of humans [47-49], One of the advancement could include the use of human bio-geographic types [44, 50] or even can be Raman spectroscopy to determine whether a used to predict certain criminal behaviors [51-53]. This suspicious powder is explosive, eliminating the need 15-dalton mark of methylation in the bases of DNA to use bleach to destroy such substances along with sequence can help to solve an array of critical cases in potential evidence [58]. Other advancement could forensic biology which when stands gloomy with include use of Fourier Transform Infrared (FTIR) current polymorphic STR technology. With the spectroscopy to identify drugs such as cocaine in advances of next-generation sequencing techniques, short span of time which could replace several heavy blooming of DNA methylation datasets and together equipments that require various gases, liquids, and with standard epigenetic protocols, the prospect of solids [59]. The use of handheld 3-D laser scanners investigating and solving these critical issues in [60, 61]and Near-infrared (Near IR) light scanners [62] forensic science is highly promising in the court of can also help scientist/police to identify a potential law. suspect during an investigation. The microfluidic Next-generation Serology chips on the other hand allow scientists to reduce contamination and build DNA profile at the scene of In very near future, mass spectrometry is going crime [5-7, 63]. Portable forensics labs could also be to be a direct method of measuring the source of equipped with devices capable of transmitting data origin of the biological forensic samples using obtained through the use of next generation facial proteomic analysis (mass spectroscopy, Electrospray recognition software and fingerprint scans to Ionization mass spectrometry (ESI-MS and Tandem government databases for comparison with stored MS) [54]. It implies that visual examination of origin files of such information. The need of the time is to of species through precipitin test (Paul Uhlenhuth Test) adopt/upgrade the forensic labs with these advanced

http://www.jbiomed.com Journal of Biomedicine 2018, Vol. 3 17 feasible technologies which could help to nab the amplification and real-time PCR in forensic casework. BMC Genomics 2009; 10: 159. perpetrators and bring justice to the innocents by 22. Valenzuela RK, Henderson MS, Walsh MH, Garrison NA, Kelch JT, Cohen-Barak O, Erickson DT, John Meaney F, Bruce Walsh J, Cheng KC, Ito S, analyzing the obscure samples which otherwise Wakamatsu K, Frudakis T, Thomas M, Brilliant MH. Predicting phenotype cannot be processed by regular instruments. This will from genotype: normal pigmentation. J Forensic Sci 2010; 55: 315-22. 23. Ambers AD, Churchill JD, King JL, Stoljarova M, Gill-King H, Assidi M, certainly help to reduce the rate of crime in future. Abu-Elmagd M, Buhmeida A, Budowle B. More comprehensive forensic genetic marker analyses for accurate human remains identification using massively parallel DNA sequencing. BMC Genomics 2016; 17: 750. Conflict of Interest 24. Draus-Barini J, Walsh S, Pospiech E, Kupiec T, Glab H, Branicki W, Kayser M. The author (Rana, A.K.) certifies that there is no Bona fide colour: DNA prediction of human eye and hair colour from ancient and contemporary skeletal remains. Investig Genet 2013; 4: 3. academic or financial interest accrued to anyone and 25. Walsh S, Kayser M. A Practical Guide to the HIrisPlex System: Simultaneous no other conflicts of interest exists whatsoever. Prediction of Eye and Hair Color from DNA. Methods Mol Biol 2016; 1420: 213-31. 26. Mazura JC, Juluru K, Chen JJ, Morgan TA, John M, Siegel EL. Facial Acknowledgements recognition software success rates for the identification of 3D surface reconstructed facial images: implications for patient privacy and security. J I sincerely thank State Forensic Science Digit Imaging 2011; 25: 347-51. 27. Broach J, Yong R, Manuell ME, Nichols C. Use of Facial Recognition Software Laboratory as well as Department of Police, State to Identify Disaster Victims With Facial Injuries. Disaster Med Public Health Government of Jharkhand, India for financial Prep 2017: 1-5. 28. Kramer KM, Hedin DS, Rolkosky DJ. Smartphone based face recognition tool assistance in the form of monthly salary for its for the blind. Conf Proc IEEE Eng Med Biol Soc 2010; 2010: 4538-41. support in bringing out this review at the State 29. Lippert C, Sabatini R, Maher MC, Kang EY, Lee S, Arikan O, Harley A, Bernal A, Garst P, Lavrenko V, Yocum K, Wong T, Zhu M, Yang WY, Chang C, Lu T, Forensic Science Laboratory Jharkhand, Ranchi, India. Lee CWH, Hicks B, Ramakrishnan S, Tang H, Xie C, Piper J, Brewerton S, Turpaz Y, Telenti A, Roby RK, Och FJ, Venter JC. Identification of individuals by trait prediction using whole-genome sequencing data. Proc Natl Acad Sci U References S A 2017. 1. Kasai K, Mukoyama H. [PCR and its application to forensic science]. 30. Hallgrimsson B, Percival CJ, Green R, Young NM, Mio W, Marcucio R. Tanpakushitsu Kakusan Koso 1990; 35: 3150-6. Morphometrics, 3D Imaging, and Craniofacial Development. Curr Top Dev 2. Kasai K, Nakamura Y, White R. Amplification of a variable number of tandem Biol 2015; 115: 561-97. repeats (VNTR) locus (pMCT118) by the polymerase chain reaction (PCR) and 31. Klingenberg CP. Size, shape, and form: concepts of allometry in geometric its application to forensic science. J Forensic Sci 1990; 35: 1196-200. morphometrics. Dev Genes Evol 2016; 226: 113-37. 3. Sgueglia JB, Geiger S, Davis J. Precision studies using the ABI prism 3100 32. Sender R, Fuchs S, Milo R. Revised Estimates for the Number of Human and genetic analyzer for forensic DNA analysis. Anal Bioanal Chem 2003; 376: Bacteria Cells in the Body. PLoS Biol 2016; 14: e1002533. 1247-54. 33. Tucker JB, Koblentz GD. The four faces of microbial forensics. Biosecur 4. Thormann W, Molteni S, Caslavska J, Schmutz A. Clinical and forensic Bioterror 2009; 7: 389-97. applications of capillary electrophoresis. Electrophoresis 1994; 15: 3-12. 34. Schmedes SE, Sajantila A, Budowle B. Expansion of Microbial Forensics. J Clin 5. Gibson-Daw G, Albani P, Gassmann M, McCord B. Rapid microfluidic Microbiol 2016; 54: 1964-74. analysis of a Y-STR multiplex for screening of forensic samples. Anal Bioanal 35. Albert J, Wahlberg J, Leitner T, Escanilla D, Uhlen M. Analysis of a rape case Chem 2017; 409: 939-47. by direct sequencing of the human immunodeficiency virus type 1 pol and gag 6. Han JP, Sun J, Wang L, Liu P, Zhuang B, Zhao L, Liu Y, Li CX. The genes. J Virol 1994; 68: 5918-24. Optimization of Electrophoresis on a Glass Microfluidic Chip and its 36. Tridico SR, Murray DC, Addison J, Kirkbride KP, Bunce M. Metagenomic Application in Forensic Science. J Forensic Sci 2017. analyses of bacteria on human hairs: a qualitative assessment for applications 7. Horsman KM, Bienvenue JM, Blasier KR, Landers JP. Forensic DNA analysis in forensic science. Investig Genet 2014; 5: 16. on microfluidic devices: a review. J Forensic Sci 2007; 52: 784-99. 37. Khodakova AS, Smith RJ, Burgoyne L, Abarno D, Linacre A. Random whole 8. Romodanovskii PO, Barinov E, Malyshev AP. [Time of death software product metagenomic sequencing for forensic discrimination of soils. PLoS One 2014; as a readily available tool for the work of a forensic medical expert at the place 9: e104996. of occurrence]. Sud Med Ekspert 2013; 56: 38-9. 38. Dirnhofer R, Jackowski C, Vock P, Potter K, Thali MJ. VIRTOPSY: minimally 9. McDown RJ, Varol C, Carvajal L, Chen L. In-Depth Analysis of Computer invasive, imaging-guided virtual autopsy. Radiographics 2006; 26: 1305-33. Memory Acquisition Software for Forensic Purposes. J Forensic Sci 2015; 61 39. Plattner T, Thali MJ, Yen K, Sonnenschein M, Stoupis C, Vock P, Suppl 1: S110-6. Zwygart-Brugger K, Kilchor T, Dirnhofer R. Virtopsy-postmortem multislice 10. Haned H, Gill P, Lohmueller K, Inman K, Rudin N. Validation of probabilistic computed tomography (MSCT) and magnetic resonance imaging (MRI) in a genotyping software for use in forensic DNA casework: Definitions and fatal scuba diving incident. J Forensic Sci 2003; 48: 1347-55. illustrations. Sci Justice 2015; 56: 104-8. 40. Thali MJ, Yen K, Schweitzer W, Vock P, Boesch C, Ozdoba C, Schroth G, Ith M, 11. Holland MM, Parson W. GeneMarker(R) HID: A reliable software tool for the Sonnenschein M, Doernhoefer T, Scheurer E, Plattner T, Dirnhofer R. Virtopsy, analysis of forensic STR data. J Forensic Sci 2010; 56: 29-35. a new imaging horizon in forensic pathology: virtual autopsy by postmortem 12. Lynch M. God's signature: DNA profiling, the new gold standard in forensic multislice computed tomography (MSCT) and magnetic resonance imaging science. Endeavour 2003; 27: 93-7. (MRI)--a feasibility study. J Forensic Sci 2003; 48: 386-403. 13. Mardis ER. Next-generation sequencing platforms. Annu Rev Anal Chem 41. Jin B, Li Y, Robertson KD. DNA methylation: superior or subordinate in the (Palo Alto Calif) 2013; 6: 287-303. epigenetic hierarchy? Genes Cancer 2011; 2: 607-17. 14. Yang Y, Xie B, Yan J. Application of next-generation sequencing technology in 42. Brunner AL, Johnson DS, Kim SW, Valouev A, Reddy TE, Neff NF, Anton E, forensic science. Genomics Proteomics Bioinformatics 2014; 12: 190-7. Medina C, Nguyen L, Chiao E, Oyolu CB, Schroth GP, Absher DM, Baker JC, 15. Melton T, Holland C, Holland M. Forensic Mitochondrial DNA Analysis: Myers RM. Distinct DNA methylation patterns characterize differentiated Current Practice and Future Potential. Forensic Sci Rev 2012; 24: 101-22. human embryonic stem cells and developing human fetal liver. Genome Res 16. Butler JM, Levin BC. Forensic applications of mitochondrial DNA. Trends 2009; 19: 1044-56. Biotechnol 1998; 16: 158-62. 43. Lokk K, Modhukur V, Rajashekar B, Martens K, Magi R, Kolde R, Koltsina M, 17. Girod A, Ramotowski R, Weyermann C. Composition of fingermark residue: a Nilsson TK, Vilo J, Salumets A, Tonisson N. DNA methylome profiling of qualitative and quantitative review. Forensic Sci Int 2012; 223: 10-24. human tissues identifies global and tissue-specific methylation patterns. 18. Wang C, Stanciu CE, Ehrhardt CJ, Yadavalli VK. Nanoscale characterization of Genome Biol 2014; 15: r54. forensically relevant epithelial cells and surface associated extracellular DNA. 44. Heyn H, Moran S, Hernando-Herraez I, Sayols S, Gomez A, Sandoval J, Monk Forensic Sci Int 2017; 277: 252-58. D, Hata K, Marques-Bonet T, Wang L, Esteller M. DNA methylation 19. Bruck S, Evers H, Heidorn F, Muller U, Kilper R, Verhoff MA. Single cells for contributes to natural human variation. Genome Res 2013; 23: 1363-72. forensic DNA analysis--from evidence material to test tube. J Forensic Sci 2011; 45. Fraga MF, Ballestar E, Paz MF, Ropero S, Setien F, Ballestar ML, Heine-Suner 56: 176-80. D, Cigudosa JC, Urioste M, Benitez J, Boix-Chornet M, Sanchez-Aguilera A, 20. Soltyszewski I, Szeremeta M, Skawronska M, Niemcunowicz-Janica A, Ling C, Carlsson E, Poulsen P, Vaag A, Stephan Z, Spector TD, Wu YZ, Plass Pepinski W. Typeability of DNA in Touch Traces Deposited on Paper and C, Esteller M. Epigenetic differences arise during the lifetime of monozygotic Optical Data Discs. Adv Clin Exp Med 2015; 24: 437-40. twins. Proc Natl Acad Sci U S A 2005; 102: 10604-9. 21. Giardina E, Pietrangeli I, Martone C, Zampatti S, Marsala P, Gabriele L, Ricci 46. Kaminsky ZA, Tang T, Wang SC, Ptak C, Oh GH, Wong AH, Feldcamp LA, O, Solla G, Asili P, Arcudi G, Spinella A, Novelli G. Whole genome Virtanen C, Halfvarson J, Tysk C, McRae AF, Visscher PM, Montgomery GW,

http://www.jbiomed.com Journal of Biomedicine 2018, Vol. 3 18

Gottesman II, Martin NG, Petronis A. DNA methylation profiles in 56. The Universal Protein Resource (UniProt) in 2010. Nucleic Acids Res 2010; monozygotic and dizygotic twins. Nat Genet 2009; 41: 240-5. 38(D): 142-8. 47. Horvath S. DNA methylation age of human tissues and cell types. Genome 57. Zhang Y, Jeltsch A. The application of next generation sequencing in DNA Biol 2013; 14: R115. methylation analysis. Genes (Basel) 2010; 1: 85-101. 48. Yi SH, Jia YS, Mei K, Yang RZ, Huang DX. Age-related DNA methylation 58. Sil S, Umapathy S. Raman spectroscopy explores molecular structural changes for forensic age-prediction. Int J Legal Med 2015; 129: 237-44. signatures of hidden materials in depth: Universal Multiple Angle Raman 49. Bekaert B, Kamalandua A, Zapico SC, Van de Voorde W, Decorte R. Improved Spectroscopy. Sci Rep 2014; 4: 5308. age determination of blood and teeth samples using a selected set of DNA 59. Marcelo MC, Mariotti KC, Ferrao MF, Ortiz RS. Profiling cocaine by methylation markers. Epigenetics 2015; 10: 922-30. ATR-FTIR. Forensic Sci Int 2014; 246: 65-71. 50. Adkins RM, Krushkal J, Tylavsky FA, Thomas F. Racial differences in 60. Polo ME, Felicisimo AM. Analysis of uncertainty and repeatability of a gene-specific DNA methylation levels are present at birth. Birth Defects Res A low-cost 3D laser scanner. Sensors (Basel) 2012; 12: 9046-54. Clin Mol Teratol 2011; 91: 728-36. 61. Hennessy RJ, McLearie S, Kinsella A, Waddington JL. Facial surface analysis 51. Szyf M. DNA methylation, behavior and early life adversity. J Genet Genomics by 3D laser scanning and geometric morphometrics in relation to sexual 2013; 40: 331-8. dimorphism in cerebral--craniofacial morphogenesis and cognitive function. J 52. Day JJ, Sweatt JD. DNA methylation and memory formation. Nat Neurosci Anat 2005; 207: 283-95. 2010; 13: 1319-23. 62. Zhang X, Bloch S, Akers W, Achilefu S. Near-infrared molecular probes for in 53. Szyf M. DNA methylation, the early-life social environment and behavioral vivo imaging. Curr Protoc Cytom 2012; Chapter 12: Unit12 27. disorders. J Neurodev Disord 2011; 3: 238-49. 63. Bruijns B, van Asten A, Tiggelaar R, Gardeniers H. Microfluidic Devices for 54. Van Steendam K, De Ceuleneer M, Dhaenens M, Van Hoofstat D, Deforce D. Forensic DNA Analysis: A Review. Biosensors (Basel) 2016; 6. Mass spectrometry-based proteomics as a tool to identify biological matrices 64. Rana, A.K., Crime investigation through DNA methylation analysis: methods in forensic science. Int J Legal Med 2012; 127: 287-98. and applications in forensics. Egypt J Forensic Sci, 2018. 8(7). 55. Allison AC, Morton JA. Species specificity in the inhibition of antiglobulin sera; a technique for the identification of human and animal bloods. J Clin Pathol 1953; 6: 314-9.

http://www.jbiomed.com